CAJ | 학술논문

目的探讨c-Jun氨基末端激酶(JNK)信号转导途径在哮喘气道重塑过程中的作用;IL-1β是否通过JNK信号途径参与哮喘气道重塑形成过程.方法 SD大鼠随机分为对照组(C)和哮喘组(A),以卵白蛋白致敏和激发复制哮喘气道重塑模型,A组根据激发时间不同,分为4、8、12周组(分别为A4、A8、A12组),同时设立相应C组(分别为C4、C8、C12组).电镜观察肺组织超微结构变化,图像分析技术测定支气管壁厚度(Wat)和平滑肌厚度(Wam);ELISA法测定血清、BALF中IL-1β 浓度;免疫组化(IHC)检测肺内磷酸化JNK(P-JNK)及其下游物磷酸化c-Jun蛋白表达;Western Blot 检测肺匀浆JNK磷酸化水平,对Wat、Wam与P-JNK蛋白平均吸光度值(mA)、P-JNK蛋白(mA)与血清、BALF IL-1β浓度进行直线相关分析.结果 4、8、12周A组War、Wam均相应地高于4、8、12周C组(均P＜0.01),12周时,A组二者均高于4周、8周(均P＜0.01);各哮喘组血清、BALF IL-1β浓度均高于同时期C组(均P＜0.01),A组BALF中IL-1β浓度,12周时,高于4周、8周(P＜0.05或P＜0.01),血清中IL-1β浓度,三者差异无统计学意义;P-JNK及其下游物P-c-Jun(mA值),各哮喘组均高于同时期c组(均P＜0.01),12周时,A组二者均高于4周、8周(均P＜0.01);Western Blot检测P-JNK蛋白吸光度值(A值),各哮喘组均高于同时期c组(均P＜0.01),A组中12周高于4周(P＜0.01),与8周组差异无统计学意义(P＞0.05);Wat、Wain与P-JNK(mA)均呈高度正相关(分别为r=0.823、r=0.818,均P＜0.01);P-JNK mA与血清、BALF IL-1β浓度均呈高度正相关(分别为r=0.717、r=0.803,均P＜0.01).结论 P-JNK及其下游物PH-Jun在哮喘大鼠气道重塑过程中表达增高,提示JNK信号通路在气道重塑进程中起重要作用;IL-1β可能部分通过激活JNK信号转导途径,参与哮喘气道重塑形成过程.
목적 탐토c-Jun안기말단격매(JNK)신호전도도경재효천기도중소과정중적작용;IL-1β시부통과JNK신호도경삼여효천기도중소형성과정.방법 SD대서수궤분위대조조(C)화효천조(A),이란백단백치민화격발복제효천기도중소모형,A조근거격발시간불동,분위4、8、12주조(분별위A4、A8、A12조),동시설립상응C조(분별위C4、C8、C12조).전경관찰폐조직초미결구변화,도상분석기술측정지기관벽후도(Wat)화평활기후도(Wam);ELISA법측정혈청、BALF중IL-1β 농도;면역조화(IHC)검측폐내린산화JNK(P-JNK)급기하유물린산화c-Jun단백표체;Western Blot 검측폐균장JNK린산화수평,대Wat、Wam여P-JNK단백평균흡광도치(mA)、P-JNK단백(mA)여혈청、BALF IL-1β농도진행직선상관분석.결과 4、8、12주A조War、Wam균상응지고우4、8、12주C조(균P＜0.01),12주시,A조이자균고우4주、8주(균P＜0.01);각효천조혈청、BALF IL-1β농도균고우동시기C조(균P＜0.01),A조BALF중IL-1β농도,12주시,고우4주、8주(P＜0.05혹P＜0.01),혈청중IL-1β농도,삼자차이무통계학의의;P-JNK급기하유물P-c-Jun(mA치),각효천조균고우동시기c조(균P＜0.01),12주시,A조이자균고우4주、8주(균P＜0.01);Western Blot검측P-JNK단백흡광도치(A치),각효천조균고우동시기c조(균P＜0.01),A조중12주고우4주(P＜0.01),여8주조차이무통계학의의(P＞0.05);Wat、Wain여P-JNK(mA)균정고도정상관(분별위r=0.823、r=0.818,균P＜0.01);P-JNK mA여혈청、BALF IL-1β농도균정고도정상관(분별위r=0.717、r=0.803,균P＜0.01).결론 P-JNK급기하유물PH-Jun재효천대서기도중소과정중표체증고,제시JNK신호통로재기도중소진정중기중요작용;IL-1β가능부분통과격활JNK신호전도도경,삼여효천기도중소형성과정.
Objective To study the role of e-Jun N-terminal kinase(JNK)signal transduction pathway in the course of asthma airway remodeling,to explore whether IL-1β participates in asthma airway remodeling mediated by JNK signal transduction pathway.Methods Totally 72 male Sprague-Dawlay rats (6-8 weeks old,weighing about 120 g)were randomly divided into control groups(36 rats)and asthma groups(36 rats).The mrs were sensitized for inducing asthma by intraperitoneal injectian of ovalbumin and AL(OH)3 and were repeatedly exposed to aeresolized ovalbumin for 4,8,12 weeks(A4,A8,or A12 group),each had 12 rats,and correspondingly control mts were intraperitoneally injected with 0.9%NaCl,then were repeatedly exposed to 0.9%NaCl for 4,8,12 weeks(CA,G8,or C12 group),each had 12 rats.The ultmstructural changes of pulmonary tissues were observed by transmission electron microscope (TEM).The total bronchial wall thickness(Wat)and the airway smooth muscle thickness(Wam)were measured byanimage analysis system.The concentrations of IL-1β in serum and broncoalveolar lavage fluid (BALF)were tested by a"sandwich"ELISA.The protein expressions of P-JNK and PH-Jun were detected by immunohistochemieal teehnique.Lung tissue extracts were analyzed for phosphorylation of JNK by Western blotting.Linear correlation analysis showed the correlation between Wat and P-JNK protein.Wam and P-JNK protein,leveh of IL-1βin serum and P-JNK protein.levels of IL-1βin BALF and P-JNK protein.Results In asthma groups,TEM showed alveolar septal proliferation and alveolus type Ⅱ epithelial cells swelling.Wat and Wam in all asthma groups were significantly higher than those in corresponding control groups(P＜0.01,respectively),and compared with group A4 and group A8,Wal and Warn ofgroup A12 significantly increased(P＜0.01).The concentrations of IL-1β in serum and BALF of asthma groups were all significantly higher than those of the corresponding control groups(P＜0.01,respectively),and compared with group A4 and group A8,the concentrations of IL-1β in BALF of group A12 significantly increased(P＜0.01 or P＜0.05).but the levels of IL-11β in serum were not significantly different among them(P＞0.05).Mean absorbance values(by immunohistochemistry)of P-JNK and e-c-Jun in asthma groups were significantly higher than those in corresponding control groups(P＜0.01,respectively),and compared with group A4 and group A8,those of group A12 significantly increased(P＜0.01 or P＜0.05).The absorbance(by Western Blot)of P-JNK in A4,A8,A12 group WaS significantly higher than that in C4,C8,C12 groups(P＜0.01,respectively),and compared with group 3.4,that of P-JNK of A12 significantly increased(P＜0.01),and compared with group A8,there was no significant difference(P＞0.05).Strong positive correlatiom were found between Wat or Wam and P-JNK(r=0.823 and r=0.818,P＜0.01,respectively,n=68)and between P-JNK and concentration of IL-1βin serum or BALF(r=0.717and,=0.803,P＜0.01,respectively,n=68).Condusions The expression of P-JNK and its downstream P-c-Jun in rats of asthma airway remodeling is increased.which implicates that JNK signal transduetion pathway plays an important role in the course of asthma airway remodeling.IL-1β participates in asthma airway remodeling possibly partly through activating JNK signal traneduction pathway.